Cloning multiple PCR products

Dag Rune Gjellesvik dagrg_nospam at online.no
Wed Mar 29 04:13:32 EST 2006


"Bibiana Silva" <bidaresonante at yahoo.com.mx> skrev i melding
news:mailman.361.1143591280.16885.methods at net.bio.net...
> Hi, everybody.
>   I have a lot of PCR bands with the same size. What can I do to separate
them?  What technique other than DGGE works better and leads to cloning and
sequencing individual products? I want to differentiate bacterial species
from metagenomic PCR amplificates.
>
>   Thanks in advance for any help.
>
>   Bibiana
>
>
Hi,
Just clone them as they are, isolate and sequence the plasmid inserts from
individual clones. Each clone will contain only one species for sure. The
chance that one cell will pick up two plasmids will be 10^-8 or something,
and you don't pick millions of colonies. But you may need a certain number
of clones to have a reasonable representation of the diversity in your
sample

Dag Rune





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